河套灌区沉积物中抗生素抗性基因的分布研究
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摘要
抗生素抗性基因(Antibiotic resistance genes, ARGs)作为一种新型污染物已经成为国内外研究的热点.为了探究河套灌区不同排水渠(3N、3S、5N、5S和7N)沉积物中ARGs的分布情况,采用定量PCR测定了沉积物中ARGs、整合酶基因(intⅠ1和intⅡ2)和转座酶基因(tnpA)的丰度.结果表明,36种ARGs中检测到31种,在各样地中均能检测到intⅠ1和tnpA基因,而intⅡ2基因未检测到.在各样地中,tetA、tetS、sulⅠ和aadA1基因丰度较高;在5N样地ereA基因丰度最高,为1.65×10~9 copies·g~(-1);15种ARGs在7N样地与其它样地之间有显著差异,其中,aadA1基因在7N样地丰度最高,为1.37×10~9 copies·g~(-1).氮含量与sulIII、ereB、bla_(ampC)基因丰度显著正相关,而pH与tetX、tetO和tetM基因丰度呈负相关关系.综上所述,抗生素抗性基因广泛存在于河套灌区的沉积物中.
Antibiotic resistance genes(ARGs), as an emerging environmental contaminant, has been a research hotspot in world. In this study, we collected sediments from different drainage channel(3 N, 3 S, 5 N, 5 S, 7 N) in Hetao irrigation district. The abundance of 36 ARGs, integrase genes(intI1 and 2) and transposase genes(tnpA) were determined quantitative real time PCR. The results showed that 31 ARGs, the intI1 and tnpA genes were detected in all the sites, but the intII2 gene was not detected. The abundances of tetA, tetS, sulI and aadA1 were high in each plot. The highest abundance ARGs was the ereA gene with 1.65×10~9 copies·g~(-1) in the 5 N plot, and aadA1 gene(1.37×10~9 copies·g~(-1)) was the highest one in the 7 N plot. The abundances of 15 ARGs in the 7 N plot were significantly different from those in other plots(p<0.05). Nitrogen demonstrated strongly positive correlations with the abundances of sulIII、ereB and bla_(ampC) genes. pH negatively correlated with the abundances of tetX, tetO, and tetM genes. In summary, ARGs are widely present in sediments in the Hetao irrigation district.
Antibiotic resistance genes(ARGs), as an emerging environmental contaminant, has been a research hotspot in world. In this study, we collected sediments from different drainage channel(3 N, 3 S, 5 N, 5 S, 7 N) in Hetao irrigation district. The abundance of 36 ARGs, integrase genes(intI1 and 2) and transposase genes(tnpA) were determined quantitative real time PCR. The results showed that 31 ARGs, the intI1 and tnpA genes were detected in all the sites, but the intII2 gene was not detected. The abundances of tetA, tetS, sulI and aadA1 were high in each plot. The highest abundance ARGs was the ereA gene with 1.65×10~9 copies·g~(-1) in the 5 N plot, and aadA1 gene(1.37×10~9 copies·g~(-1)) was the highest one in the 7 N plot. The abundances of 15 ARGs in the 7 N plot were significantly different from those in other plots(p<0.05). Nitrogen demonstrated strongly positive correlations with the abundances of sulIII、ereB and bla_(ampC) genes. pH negatively correlated with the abundances of tetX, tetO, and tetM genes. In summary, ARGs are widely present in sediments in the Hetao irrigation district.
引文
陈超琼.2017.抗生素在中国东南沿海沉积物中的分布初探[D].上海:华东师范大学
Chen B W, Liang X M, Huang X P, et al.2013.Differentiating anthropogenic impacts on ARGs in the Pearl River Estuary by using suitable gene indicators[J].Water Research,47(8):2811-2820
Crecchio C,Ruggiero P,Curci M,et al.2005.Binding of DNA from Bacillus subtilis on montmorillonite-humic acids-aluminum or iron hydroxypolymers:Effects on transformation and protection against DNase[J].Soil Science Society of America Journal,69(3):834-841
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贾励瑛.2014.β-内酰胺类抗生素在污水处理厂的分布与迁移转化[D].邯郸:河北工程大学
江月,陈奕涵,何义亮.2018.长江下游某水源型水库抗生素抗性基因污染研究[J].环境科学学报,38(3):884-891
Jiang L,Hu X,Xu T,et al.2013.Prevalence of antibiotic resistance genes and their relationship with antibiotics in the Huangpu River and the drinking water sources,Shanghai,China[J].Science of the Total Environment,458:267-272
Jiang L,Hu X L,Xu T,et al.2013.Prevalence of antibiotic resistance genes and their relationship with antibiotics in the Huangpu River and the drinking water sources,Shanghai,China[J].Science of the Total Environment,4(38):267-272
Li C,Lu J J,Liu J,et al.2016.Exploring the correlations between antibiotics and antibiotic resistance genes in the wastewater treatment plants of hospitals in Xinjiang,China[J] Environmental Science and Pollution Research,23(15):15111-15121
Li J J,Xin Z H,Zhang Y Z,et al.2017.Long-term manure application increased the levels of antibiotics and antibiotic resistance genes in a greenhouse soil[J].Applied Soil Ecology,121:193-200
Luan X B,Wu P T,Sun S K,et al.2018.Impact of land use change on hydrologic processes in a large plain irrigation district[J].Water Resources Management,32(9):3203-3217
罗义,周启星.2008.抗生素抗性基因(ARGs):一种新型环境污染物[J].环境科学学报,28(8):1499-1505
Luo Y,Mao D Q,Rysz M,et al.2010 Trends in antibiotic resistance genes occurrence in the Haihe River,China[J].Environmental Science and Technology,44(19):7220-7225
Luo Y,Xu L,Rysz M,et al.2011.Occurrence and transport of tetracycline,sulfonamide,quinolone,and macrolide antibiotics in the Haihe River Basin,China[J].Environmental Science and Technology,45(5):1827-1833
Marti E,Variatza E,Balcazar J L.2014.The role of aquatic ecosystems as reservoirs of antibiotic resistance[J].Trends in Microbiology,22(1):36-41
Martinez J L.2009.Environmental pollution by antibiotics and by antibiotic resistance determinants[J].Environmental Pollution,157(11):2893-2902
Martínez J L.2008.Antibiotics and antibiotic resistance genes in natural environments[J].Science,321(5887):365-367
Ouyang W Y,Huang F Y,Zhao Y,et al.2015.Increased levels of antibiotic resistance in urban stream of Jiulongjiang River,China[J] Applied Microbiology and Biotechnology,99(13):5697-5707
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Rahube T O,Marti R,Scott A,et al.2016.Persistence of antibiotic resistance and plasmid-associated genes in soil following application of sewage sludge and abundance on vegetables at harvest[J].Canadian Journal of Microbiology,62(7):600-607
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Schwartz T,Kohnen W,Jansen B,et al.2003.Detection of antibiotic-resistant bacteria and their resistance genes in wastewater,surface water,and drinking water biofilms[J].FEMS Microbiology Ecology,43(3):325-335
Stoll C,Sidhu J P S,Tiehm A,et al.2012.Prevalence of clinically relevant antibiotic resistance genes in surface water samples collected from Germany and Australia[J].Environmental Science and Technology,46(17):9716-9726
Szekeres E,Chiriac C M,Baricz A,et al.2018.Investigating antibiotics,antibiotic resistance genes,and microbial contaminants in groundwater in relation to the proximity of urban areas[J].Environmental Pollution,236:734-744
Wang F H,Qiao M,Chen Z,et al.2015.Antibiotic resistance genes in manure- amended soil and vegetables at harvest[J].Journal of Hazardous Materials,299:215-221
Wang F H,Qiao M,Lv Z E,et al.2014.Impact of reclaimed water irrigation on antibiotic resistance in public parks,Beijing,China[J].Environmental Pollution,184:247-253
徐晨.2017.三峡库区水体、土壤和沉积物中抗生素与抗生素抗性基因的污染特征[D].武汉:中国科学院武汉植物园
Xu Y,Guo C S,Luo Y,et al.2016.Occurrence and distribution of antibiotics,antibiotic resistance genes in the urban rivers in Beijing,China[J].Environmental Pollution,213:833-840
Yang Y,Xu C,Cao X,et al.2017a.Antibiotic resistance genes in surface water of eutrophic urban lakes are related to heavy metals,antibiotics,lake morphology and anthropic impact[J].Ecotoxicology,26(6):831-840
Yang Y Y,Liu W Z,Xu C,et al.2017b.Antibiotic resistance genes in lakes from middle and lower reaches of the Yangtze River,China:effect of land use and sediment characteristics[J].Chemosphere,178:19-25
张红,程寒飞.2018.水环境中氨基糖苷类抗性基因污染及研究进展[J].环境科学与技术,41(10):1-10
张宏杰,2011.海河流域中抗生素抗性基因分布及有关抗性基因的进化分析[D].沈阳:沈阳药科大学
赵溪,李君剑,李洪建.2010.关帝山不同植被恢复类型对土壤碳、氮含量及微生物数量的影响[J].生态学杂志,29(11):2102-2110
Zhou L,Ying G,Liu S,et al.2013.Excretion masses and environmental occurrence of antibiotics in typical swine and dairy cattle farms in China[J].Science of the Total Environment,444:183-195
Zhu Y G,Johnson T A,Su J Q,et al.2013.Diverse and abundant antibiotic resistance genes in Chinese swine farms[J].Proceedings of the National Academy of Sciences of the United States of America,110(9):3435-3440
Zhu Y G,Johnson T A,Su J Q.et al.2013.Diverse and abundant antibiotic resistance genes in Chinese swine farms[J].PNAS,110(9):3435-3440
Chen B W, Liang X M, Huang X P, et al.2013.Differentiating anthropogenic impacts on ARGs in the Pearl River Estuary by using suitable gene indicators[J].Water Research,47(8):2811-2820
Crecchio C,Ruggiero P,Curci M,et al.2005.Binding of DNA from Bacillus subtilis on montmorillonite-humic acids-aluminum or iron hydroxypolymers:Effects on transformation and protection against DNase[J].Soil Science Society of America Journal,69(3):834-841
冯兆忠,王效科,冯宗炜,等.2003.河套灌区秋浇对不同类型农田土壤氮素淋失的影响[J].生态学报,23(10):2027-2032
Hill K E,Top E M.1998.Gene transfer in soil systems using microcosms[J].FEMS Microbiology Ecology,25(4):319-329
贾励瑛.2014.β-内酰胺类抗生素在污水处理厂的分布与迁移转化[D].邯郸:河北工程大学
江月,陈奕涵,何义亮.2018.长江下游某水源型水库抗生素抗性基因污染研究[J].环境科学学报,38(3):884-891
Jiang L,Hu X,Xu T,et al.2013.Prevalence of antibiotic resistance genes and their relationship with antibiotics in the Huangpu River and the drinking water sources,Shanghai,China[J].Science of the Total Environment,458:267-272
Jiang L,Hu X L,Xu T,et al.2013.Prevalence of antibiotic resistance genes and their relationship with antibiotics in the Huangpu River and the drinking water sources,Shanghai,China[J].Science of the Total Environment,4(38):267-272
Li C,Lu J J,Liu J,et al.2016.Exploring the correlations between antibiotics and antibiotic resistance genes in the wastewater treatment plants of hospitals in Xinjiang,China[J] Environmental Science and Pollution Research,23(15):15111-15121
Li J J,Xin Z H,Zhang Y Z,et al.2017.Long-term manure application increased the levels of antibiotics and antibiotic resistance genes in a greenhouse soil[J].Applied Soil Ecology,121:193-200
Luan X B,Wu P T,Sun S K,et al.2018.Impact of land use change on hydrologic processes in a large plain irrigation district[J].Water Resources Management,32(9):3203-3217
罗义,周启星.2008.抗生素抗性基因(ARGs):一种新型环境污染物[J].环境科学学报,28(8):1499-1505
Luo Y,Mao D Q,Rysz M,et al.2010 Trends in antibiotic resistance genes occurrence in the Haihe River,China[J].Environmental Science and Technology,44(19):7220-7225
Luo Y,Xu L,Rysz M,et al.2011.Occurrence and transport of tetracycline,sulfonamide,quinolone,and macrolide antibiotics in the Haihe River Basin,China[J].Environmental Science and Technology,45(5):1827-1833
Marti E,Variatza E,Balcazar J L.2014.The role of aquatic ecosystems as reservoirs of antibiotic resistance[J].Trends in Microbiology,22(1):36-41
Martinez J L.2009.Environmental pollution by antibiotics and by antibiotic resistance determinants[J].Environmental Pollution,157(11):2893-2902
Martínez J L.2008.Antibiotics and antibiotic resistance genes in natural environments[J].Science,321(5887):365-367
Ouyang W Y,Huang F Y,Zhao Y,et al.2015.Increased levels of antibiotic resistance in urban stream of Jiulongjiang River,China[J] Applied Microbiology and Biotechnology,99(13):5697-5707
Pruden A,Arabi M,Storteboom H N.2012.Correlation between upstream human activities and riverine antibiotic resistance genes[J].Environmental Science and Technology,46(21):11541-11549
Pruden A,Pei R,Storteboom H,et al.2006.Antibiotic resistance genes as emerging contaminants:Studies in Northern Colorado?[J].Environment Science and Technology,40(23):7445-7450
Rahube T O,Marti R,Scott A,et al.2016.Persistence of antibiotic resistance and plasmid-associated genes in soil following application of sewage sludge and abundance on vegetables at harvest[J].Canadian Journal of Microbiology,62(7):600-607
沈群辉,冀秀玲,傅淑珺,等.2012.黄浦江水域抗生素及抗性基因污染初步研究[J].生态环境学报,21(10):1717-1723
Schwartz T,Kohnen W,Jansen B,et al.2003.Detection of antibiotic-resistant bacteria and their resistance genes in wastewater,surface water,and drinking water biofilms[J].FEMS Microbiology Ecology,43(3):325-335
Stoll C,Sidhu J P S,Tiehm A,et al.2012.Prevalence of clinically relevant antibiotic resistance genes in surface water samples collected from Germany and Australia[J].Environmental Science and Technology,46(17):9716-9726
Szekeres E,Chiriac C M,Baricz A,et al.2018.Investigating antibiotics,antibiotic resistance genes,and microbial contaminants in groundwater in relation to the proximity of urban areas[J].Environmental Pollution,236:734-744
Wang F H,Qiao M,Chen Z,et al.2015.Antibiotic resistance genes in manure- amended soil and vegetables at harvest[J].Journal of Hazardous Materials,299:215-221
Wang F H,Qiao M,Lv Z E,et al.2014.Impact of reclaimed water irrigation on antibiotic resistance in public parks,Beijing,China[J].Environmental Pollution,184:247-253
徐晨.2017.三峡库区水体、土壤和沉积物中抗生素与抗生素抗性基因的污染特征[D].武汉:中国科学院武汉植物园
Xu Y,Guo C S,Luo Y,et al.2016.Occurrence and distribution of antibiotics,antibiotic resistance genes in the urban rivers in Beijing,China[J].Environmental Pollution,213:833-840
Yang Y,Xu C,Cao X,et al.2017a.Antibiotic resistance genes in surface water of eutrophic urban lakes are related to heavy metals,antibiotics,lake morphology and anthropic impact[J].Ecotoxicology,26(6):831-840
Yang Y Y,Liu W Z,Xu C,et al.2017b.Antibiotic resistance genes in lakes from middle and lower reaches of the Yangtze River,China:effect of land use and sediment characteristics[J].Chemosphere,178:19-25
张红,程寒飞.2018.水环境中氨基糖苷类抗性基因污染及研究进展[J].环境科学与技术,41(10):1-10
张宏杰,2011.海河流域中抗生素抗性基因分布及有关抗性基因的进化分析[D].沈阳:沈阳药科大学
赵溪,李君剑,李洪建.2010.关帝山不同植被恢复类型对土壤碳、氮含量及微生物数量的影响[J].生态学杂志,29(11):2102-2110
Zhou L,Ying G,Liu S,et al.2013.Excretion masses and environmental occurrence of antibiotics in typical swine and dairy cattle farms in China[J].Science of the Total Environment,444:183-195
Zhu Y G,Johnson T A,Su J Q,et al.2013.Diverse and abundant antibiotic resistance genes in Chinese swine farms[J].Proceedings of the National Academy of Sciences of the United States of America,110(9):3435-3440
Zhu Y G,Johnson T A,Su J Q.et al.2013.Diverse and abundant antibiotic resistance genes in Chinese swine farms[J].PNAS,110(9):3435-3440